Effects of sodium butyrate on proliferation-dependent insulin gene expression and insulin release in glucose-sensitive RIN-5AH cells

• Published in: The Journal of biological chemistry 1991-04, Vol.266 (12), p.7542-7548
• Main Authors: Karlsen, A E, Fujimoto, W Y, Rabinovitch, P, Dube, S, Lernmark, A
• Format: Article
• Language: English
• Subjects: Animals; Biological and medical sciences; Blotting, Northern; Butyrates - pharmacology; Butyric Acid; Cell Cycle; Fundamental and applied biological sciences. Psychology; Gene expression; Gene Expression Regulation - drug effects; Glucose - pharmacology; Insulin - genetics; Insulin - metabolism; Insulin Secretion; Molecular and cellular biology; Molecular genetics; Nucleic Acid Hybridization; Radioimmunoassay; Rats; RNA - analysis; Tumor Cells, Cultured - pathology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(20)89481-2

A rat islet tumor subclone, RIN-5AH-T2-B, was cultured with 2 mmol/liter of the proliferation-arresting compound sodium butyrate (NaB). Insulin gene expression and glucose-stimulated insulin release were analyzed and compared with logarithmically proliferating and confluent control cells cultured without NaB. Logarithmically proliferating control cells revealed high insulin gene expression. In the presence of amino acids, these cells showed a dose-dependent insulin response to glucose with a half-maximal and maximal 6.5-fold stimulation by 0.8 and 5.6 mmol/liter D-glucose, respectively. However, as the control cells approached growth arrest, insulin gene expression subsided to below detectability, an occurrence that is associated with decreased insulin release and accumulation of cells in the G1 phase of the cell cycle. In contrast, NaB-arrested cells showed continuous insulin gene expression throughout the experiment. Despite this, insulin release in response to glucose was lost. NaB revealed a biphasic effect on the cell-cycle: after an initial leaky G1 arrest during the first 24 h, the 5AH-B cells were arrested in G2 during the following 3 days. These data suggest that insulin gene expression and glucose-stimulated insulin release are affected by the cell cycle. These glucose-sensitive RIN-5AH-T2-B cells may be useful in studies of insulin secretion and gene regulation.